Dissecting the mechanics of alloproliferation with single cell RNA-seq

January 10, 2016

Central Dogma of T cell activation may be too simplistic

PBMCS from two donors, irradiate one fraction and label other
Mix together
Allow alloactivation of T cells to proceed over the following 3-5 days
Dividing T cells will dilute out the green label (CFSE) and can be quantified flow cytometrically

<<<<<<< HEAD How can single cell RNA seq performed on cells from an MLR inform us of the transcriptional events that define alloproliferation?

======= How can single cell RNA seq performed on cells from an MLR inform us of the transcriptional events that define alloproliferation

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Three donors were utilized (A, B, C)
MLR was set up using each donor as a responder on each other donor (e.g A on B, A on C, B on A, B on C)
Controls included T cells cultured with anti-CD3/anti-CD28 microbeads and T cells cultures with autologous controls
Cells were harvested and sorted by CFSE dilution (e.g. Division 0, Division 1, … Division 4)
Cells were droplet partitioned and mRNA libraries created using the 10X genomics platform

The end result